Generation of the panel of transgenic human cell lines with stable expression of mutant variants of the GJB2 gene associated with hearing loss for comparative in vitro studies

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Ekaterina Alexandrovna Maslova¹, Marina Vyacheslavovna Zytsar², Valeriia Yuryevna Danilchenko³, Olga Leonidovna Posukh⁴, Konstantin Evgenyevich Orishchenko^5
1Federal Research Center Institute of Cytology and Genetics, Novosibirsk State University, Novosibirsk, Russian Federation, maslova@bionet.nsc.ru
²Federal Research Center Institute of Cytology and Genetics, Novosibirsk, Russian Federation, zytzar@bionet.nsc.ru
³Federal Research Center Institute of Cytology and Genetics, Novosibirsk, Russian Federation, danilchenko_valeri@mail.ru
⁴Federal Research Center Institute of Cytology and Genetics, Novosibirsk State University, Novosibirsk, Russian Federation, posukh@bionet.nsc.ru
⁵Federal Research Center Institute of Cytology and Genetics, Novosibirsk State University, Novosibirsk, Russian Federation, OrishchenkoKE@icg.sbras.ru

Assessment of pathogenicity of novel variants is a primary task for molecular diagnostics of hereditary diseases, and comprehensive evidences, including functional in vitro studies, are necessary to support their involving in pathology. Mutations in the GJB2 gene encoding transmembrane protein Cx26 are the common cause for hearing loss worldwide. About 400 pathogenic and also not yet classified variants with ambiguous functional significance are described in the GJB2 gene sequence. In this study, for the first time, the panel of transgenic HeLa cell lines stably expressing of different GJB2 variants was generated for subsequent comparative analysis of structure and functioning of the Cx26 protein. This panel was successfully applied for confirmation of the pathogenicity of novel GJB2 missense variant c.516G>C (p.Trp172Cys) found with high frequency in deaf patients from Southern Siberia.